A protein-acetylenic ester complex to retard digestion in the rumen

ABSTRACT

Improved protein feed material for ruminants which is resistant to digestive breakdown in the rumen but not in the abomasum and/or intestines which comprises the reaction product of a protein-containing feed material and an ester selected from the group consisting of acetylenic monoesters and acetylenic diesters. Exemplary of such acetylenic esters are ethylene glycol, bispropiolate and n-decyl propiolate.

United States PHtEIlt 91 Wildi et a1.

[ 51 Feb. 27, 1973 A PROTEIN-ACETYLENIC ESTER COMPLEX TO RETARD DIGESTION IN THE RUMEN [75] Inventors: Bernard S. Wildi, Kirkwood; Robert E. Miller, Ballwin, all of Mo.

[73] Assignee: Monsanto Company, St. Louis, Mo.

[22] Filed: Oct. 27, 1970 [21] Appl. No.: 84,478

[52] US. Cl ..99/2 R, 106/124, 106/138, 106/154 R, 260/112 R [51] Int. Cl. ..A23k 1/18 [58] Field of Search ...99/2 R, 14, 17, 18; 260/112 R, 260/112 G,117,119,123,123.5;8/94.33; 106/124,l38, 154 R [56] References Cited UNITED STATES PATENTS 3,082,245 3/1963 Miller ..260/486 R OTHER PUBLICATIONS Chem. Abstracts, Vol. 43, par. 7113 (i), 1949 Lennox, F.G.

Chem. Abstracts, Vol. 44, par. 193 (a), 1950 E. A. Prill et a].

Chem. Abstracts, Vol. 52, par. 10186 (f), 1958 H. M. Teeter et a1.

Chem. Abstracts, Vol. 41, par 2195 (d), 1947 P. W. Brian at a].

Primary Examiner-Norman Yudkoff Assistant Examiner-Kenneth Van Wyck Attorney-Lynden N. Goodwin, Neal E. Willis and James W. Williams, Jr.

57 I ABSTRACT 5 Claims, No Drawings A PROTElN-ACETYLENIC ESTER COMPLEX TO RETARD DIGESTION IN THE RUMEN BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates to a method for improving the feed utilization of ruminant animals. In a particular aspect this invention relates to a method for improving protein utilizationin ruminant animals. In a further aspect this invention relates to modified protein feed compositions useful in ruminant nutrition which are resistant to digestive attack in the fluid medium of the rumen.

2. Description of the Prior Art The digestive system of the ruminant animal (cattle, sheep, bison, camels, etc.) is designed to permit efficient use of coarse, fibrous foodstuffs. Because of its particular structure and nature, however, the ruminants digestive system is inefficient in obtaining nutritional value from protein materials. Principally for this reason it is common practice in ruminant nutrition to supplement the diet of the animal with added protein. The supplemental protein serves to increase the rate of growth of the animal and in the case of sheep promotes wool growth.

The rumen, the largest of the four stomach compartments of the animal, serves as an important location for digestive breakdown of ingested foodstuffs chiefly through the action of microorganisms present therein. However, absorption of most nutrients for metabolic purposes does not occur in the rumen but takes place further along in the alimentary tract, principally in the abomasum and intestines. ingested food is typically retained in the rumen for from about 12-30 hours during which time it is subject to digestive breakdown by the microorganisms and by the rumen fluid. Much ingested protein material is broken down in the rumen to soluble peptides and amino acids. In turn much of these peptides and amino acids are utilized by the microorganisms present in the rumen fluid thereby removing them as a source of nutrition for the host animal.

Because of the desirability as indicated above of avoiding protein breakdown in the rumen-in order to permit absorption in the abomasum and intestines it has been suggested that nutrient protein containing materials fed to ruminants be treated so as to permit passage without digestive breakdown through the rumen to the abomasum. Suggested procedures have included coating the protein material, for example, with fats and vegetable oils, heat treatment of the protein material and reaction of the protein material with formaldehyde. In any event the treated material must be resistant to digestive breakdown in the rumen fluid, which is a fluid buffered at about pI'I6-7 by phosphate-bicarbonate from saliva and carbon dioxide, but subject to breakdown in the acid medium of the fluid of the abomasum which has a pH, due principally to hydrochloric acid secretion, of about 2-4.

OBJECTS It is an object of the present invention to provide a method for improving the feed utilization of ruminant animals.

It is a further object of the present invention to provide a method for improving the protein utilization of ruminant animals whereby protein passes through the rumen without substantial digestive breakdown.

Other objects and advantages of the present invention will be apparent from the specification and appended claims.

SUMMARY OF THE INVENTION It has been found in accordance with the present invention that the protein utilization of the ruminant animal is improved by feeding the animal a reaction prdduct of a protein-containing nutrient material and an ester which is an acetylenic monoester or an acetylenic diester.

The amount of acetylenic ester in the reaction product is sufficient to prevent substantial digestive breakdown in the fluid medium of the rumen but insufficient to prevent digestive breakdown in the fluid media of the abomasum and of the intestines.

DETAILED DESCRIPTION The protein-containing reaction product used in the method of the. present invention is the reaction product of a protein-containing nutrient feed material and an acetylenic ester. Acetylenic esters suitable for use in the process of the present invention are represented by the following general formula wherein Z is hydrogen, lower alkyl, for example, 1-5 carbon atoms or aryl, X is R or the radical R is alkyl, for example, l-18 carbon atoms or aryl and R is a divalent hydrocarbon radical, for example, alkylene having 1-l8 carbon atoms or arylene, for example, phenylene. Suitable acetylenic esters include 2-butene-l,4-diol dipropiolate, ethylene glycol ditetrolate, 2,3-dimethyl-l,3-pentane diol di-2-pentynoate, propylene glycol dipropiolate, 2-methyl-l,5-pentane diol bis(phenyl propiolate), 2-heptene-l,6-diol dipropiolate, l,6-dimethyl-l,2-heptadecane diol di-2- pentynoate, hydroquinone dipropiolate, benzene diethanol bis(phenyl I propiolate), l-phenyl-l,2'- propane diol dipropiolate, p,p'-dicyclohexanol dipropiolate, 1,2-cyclopentane diol bis(phenyl propiolate), 4-cyclohexene-l,Z-dimethanol dl-Z-pentynoate, 1,1-cyclo propane dimethanol dipropiolate, cyclohexane-l,2-ethane diol bis(phenyl propiolate), 2-methyl- 1,2-propane diol dipropiolate, 2-methyl-2-octene-l,4- diol dipropiolate, 4,4'-diphenol bispropiolate, 2,2- dichloromethyl-l,S-propane diol bispropiolate, methyl propiolate, n-decyl propiolate, phenyl propiolate, etc.

The protein-containing nutrient material can be from any suitable source including animal, plant, or synthetic sources such as, for example, silage, grains, nuts, chalfs, casein, soy bean meal, fish meal, peanut meal, beef scraps, pork scraps, linseed meal, milk solids, etc.

The useful reaction products of the present invention are prepared by the interaction of acetylenic ester and protein nutrient material by any suitable procedure. The reaction is readily carried out in a suitable solvent medium inert to the reactants and the reaction products in the presence of a basic catalyst, for exampie, an alkali metal hydroxide, an alkali metal acetate, or a quaternary ammonium salt. A pH range for reaction of 9-11 is preferred. Inert solvents suitable for use include water and lower alcohols such as ethanol and methanol. The reaction may be carried out at any suitable temperature, however, elevated temperatures above about 70C particularly for extended periods should be avoided to minimize degradation of protein material. Temperatures in the range of from about 10 to about 40C are typically employed with room temperature being both suitable and practical. The reaction is preferably carried out simply by forming a slurry or solution of the reactants in the solvent medium and agitating, as by stirring, the slurry thereby to permit sufficient reaction of the protein with the acetylenic ester. The thus treated protein is then recovered by filtration, or by precipitation and filtration followed by drying as by oven drying, drum drying, or simple evaporation to obtain the modified protein nutrient material.

It is important in order to insure operability of the present invention that the amount of acetylenic ester incorporated into the protein be sufficient to prevent digestive breakdown to soluble peptides and amino acids in the rumen but insufficient to prevent digestive breakdown to soluble peptides and amino acids in the abomasum and intestines. This amount will, of course, vary and will depend among other things on the particular protein material, the particular acetylenic ester of choice, the pH of the solvent of reaction, time and temperature of reaction, the species and age of the animal, and the total makeup of the animal diet. Typically an amount in the range of from about 0.0001 to about 0.01 mole of acetylenic ester for each gram of protein contained in the protein material is employed. When the acetylenic ester is a monoester an amount in the range of from about 0.0005 to about 0.002 mole per gram of protein is preferred whereas when the acetylenic ester is a diester an amount in the range of from about 0.0005 to about 0.006 mole is preferred.

It is to be understood that the modified ruminant protein feed of the present invention can be fed separately to the animal or it can be used for incorporation in other ruminant feed materials. Illustrative of ruminant feed materials in which the protein material of the present invention may be incorporated are soy bean meal, ground corn, hay, straw, cotton seed hulls, cotton mill waste, feed pulp, silage, oats, barley, cereal, brans, cereal middlings and combinations thereof. If desired other components, for example, minerals, such as bone meal, salt, and trace minerals, antibiotics and vitamins may be included in the animal feed ration.

The following examples illustrate the effectiveness of acetylenic ester compositions useful in the present invention in protecting protein-containing material from digestion in the fluid of the rumen while permitting digestion in the fluids of the abomasum and intestines. The small scale in vitro experiments shown in the examples simulate conditions existing in the rumen, in the abomasum and in the intestines thereby permitting the study of treated protein without the use of the live animal and large quantities of feed materials. It is understood that the examples are presented for the purpose of illustration only and the invention is not limited to the compositions or methods shown therein.

EXAMPLE 1 Preparation of Treated Protein Casein (5.01 grams) was dissolved in a solution of ethyiene glycol bispropiolate (0.61 gram) in aqueous sodium hydroxide (pHlO-l l) (60 milliliters). The resulting mixture was stirred for about 6-8 hours at room temperature to permit reaction of the ethylene glycol bispropiolate with the casein. The reaction product was isolated by acidification to pl-l3.5-3.8, filtered, washed with water and vacuum dried to obtain protein product containing 0.0007 mole of ethylene glycol bispropiolate per gram of casein.

Rumen Digestion Test To 10 milliliters of rumen fluid from fasted sheep contained in a 50 milliliter glass flask was added 10 milliliters of a buffered solution of the following composition.

BUFFER SOLUTION IN GRAMS PER LITER NaH,PO 0.3l6 xmro. 0.152 NaHCO, 2.260 KCl 0.375 M so. 0.112 NaCl 0.375 cacl, 0.03s Fes0.-7H,o 0.00s Mnso. 0.004 znso,-7H,0 0.004 cus0.-sH,o 0.002 cocl, 0.001

The resulting mixture was adjusted to pl-l6.8 (4 N l-lCl). To the buffered rumen fluid was added milligrams of the ethylene glycol bispropiolate treated protein prepared above. The flask was then purged with nitrogen, stoppered (pressure release valve) and heated at 38C on a water shaker bath. Protection of protein from digestion was determined by ammonia production with a lower amount of ammonia production indicating a lower amount of digestion of protein. Ammonia production was determined after 6 hours and after 24 hours with results being presented in Table 1.

Abomasum Digestion Test Gastric fluid was prepared as follows: NaCl (2 grams) was dissolved in sufficient water to give a total volume of 950 milliliters. Pepsin (3.2 grams) was added thereto. Concentrated hydrochloric acid (7 milliliters) was added to the resulting medium and the pH of the medium was then adjusted to 2.0 with aqueous sodium hydroxide.

To a glass flask containing 20 milliliters of the gastric fluid were added 60 milligrams of ethylene glycol bispropiolate treated casein prepared above. The glass flask containing the resulting mixture was stoppered with pressure release valves and heated at 38C on a water shaker bath for 2 hours. Digestion of protein was then determined by ammonia analysis, the greater amount of ammonia produced the greater the amount of protein digested. The results are given in Table l.

Intestine Digestion Test Intestinal fluid was prepared as follows: NaCl (2 grams) was dissolved in sufficient water to give a total volume of 950 milliliters. Pepsin (3.2 grams) was added thereto. Concentrated hydrochloric acid (7 milliliters) was added to the medium and the pH of the medium was then adjusted to 7.0 with 0.1 N sodium hydroxide.

' 5 6 Pancreatin (10 milligrams per milliliter of medium) was added to the resulting medium. Z, CEC C. Q X

To a glass flask containing 20 milliliters of the intestinal fluid were added 60 milligrams of ethylene glycol bispropiolate treated casein prepared above. v V The glass flask was stoppered with pressure release wherein Z is selected from the group consisting of valves and heated at 38C on a water shaker bath for hydrogen, lower alkyl radical having 1-5 carbon atoms the prescribed period of time. Digestion of protein was and aryl radical, and X is selected from the group condetermined by ammonia analysis, the greater amount sisting of R and the radical of ammonia produced the greater amount of protein digested. The results are given in Table 1. 0

EXAMPLES 2-9 eel-0a Following the general procedures and tests of Example l acetylenic ester treated casein samples were wherein R is selected from the group consisting of alkyl prepared and tested. The results are given in Table l. radical having l-18 carbon atoms and aryl radical and In the same manner other protein-containing materi- R' is a divalent hydrocarbon radical selected from the als may be reacted with acetylenic esters to protect the group consisting of alkylene having 1-18 carbon atoms protein material from digestion in the rumen while perand arylene, the amount of acetylenic ester in said mitting its digestion in the abomasum and intestines. 20 complex being in the range of from about 0.0001 to Since many embodiments of this invention may be about 0.01 mole per gram of protein, said amount made and since many changes may be made in the embeing sufficicnt to render said feed material resistant to bodiments described, the foregoing is to be interpreted digestive breakdown in the fluid of the rumen but insufas illustrative only and the invention is defined by the ficient to prevent substantial digestive breakdown in claims appended hereto. the fluids of the abomasum and of the intestines.

TABLE 1 Percent total protein digested Mole Ammonia N Ammonia N Ammonia N ester] rumen abomesum intestine Example Solvent oi gram number Aeotylenieostor preparation casein 0hrs. 24hrs. Zhrs. ihrs. 20hrs.

l. Ethylene glycol blspr0pi0late.... Aqueous NaOH..... 0.0007 0 5.7 32 38 54 2 .00"... ..do 0.0003 "8 25 0 81) 100 3 110.. do 0. 001 40.5 3.1 24 37 50 4, d0 Ethanol (aqueous). 0.0001 64.6 62.3 05 107 113 5 4,4- phenol bispropiolnto do 0.0037 28.7 10.8 39.4 0 do do... 0.0007 0 1.2 43.0 10 0s 7 n-Decyl propiolate d0.. O 0013 '39 18 42 60 8 Methyl propiolate d0. 0.0006 70 61 83 03 115 0 d0 do.-.. 000020 *100 100 10 82 100 Due at least in part to decomposition of rumen microorganisms.

We claim: 2. The method of claim 1 wherein the acetylenic l. A method for improving the protein utilization of ester is an acetylenic monoester. ruminant animals which comprises feeding the rumi- 3. The method of claim 2 wherein the acetylenic nant animal a complex of a protein-containing nutrient monoester is n-decyl propiolate. feed material selected from the group consisting of 4. The method of claim 1 wherein the acetylenic silage, grain, nuts, chaffs, caseins, soybean meal, fish 45 ester isanacetylenic diester. meal, peanut meal, beef scraps, pork scraps, linseed 5. The method of claim 4 wherein the acetylenic dimeal and milk solids and an acetylenic ester of the forester is ethylene glycol bispropiolatc. mula t =0 e 

2. The method of claim 1 wherein the acetylenic ester is an acetylenic monoester.
 3. The method of claim 2 wherein the acetylenic monoester is n-decyl propiolate.
 4. The method of claim 1 wherein the acetylenic ester is an acetylenic diester.
 5. The method of claim 4 wherein the acetylenic diester is ethylene glycol bispropiolate. 